Lidocaine inhibits NIH-3T3 cell multiplication by increasing the expression of cyclin-dependent kinase inhibitor 1A (p21).
BACKGROUND
We explored molecular mechanisms by which lidocaine inhibits growth in the murine embryonic fibroblast cell line NIH-3T3. Local anesthetics can adversely affect cell growth in vitro. Their effects on wound healing are controversial. We examined the effects and novel mechanisms by which lidocaine affects in vitro multiplication of the murine fibroblast cell line NIH-3T3.


METHODS
NIH-3T3 cells were grown in culture with lidocaine [0, 0.05, 0.5, 1, 2, and 5 mM]. Cell multiplication was assessed by determining cell counts on subsequent days, while mechanisms by which inhibition occurred were evaluated by bromodeoxyuridine uptake, gene expression using polymerase chain reaction array, and Western blot analysis to verify increased levels of affected proteins.


RESULTS
Lidocaine caused dose-dependent inhibition of multiplication of NIH-3T3 cells. Effects ranged from no inhibition [0.05 and 0.5 mM] and mild inhibition [1 mM], to severe inhibition [2 and 5 mM] [P = 0.006]. Lidocaine 2 mM inhibited bromodeoxyuridine uptake at day 3.5 [P = 0.02 versus control, and P = 0.0495 vs 1 mM lidocaine]. On day 1.5, lidocaine upregulated expression of cyclin-D1 and cyclin-dependent kinase inhibitor 1A [p21]. On day 2.5, lidocaine increased the levels of p21 protein.


CONCLUSIONS
Low concentrations of lidocaine, as would be seen in plasma after spinal, epidural, or plexus anesthesia, do not significantly affect multiplication of fibroblasts. Higher doses of lidocaine arrest cell multiplication at the S-phase of the growth cycle by upregulation of p21, an extremely potent inhibitor of cell multiplication. Higher concentrations, as would be seen after tissue infiltration, severely inhibit fibroblast multiplication and thus may impair wound healing.
Desai
Sukumar P
SP
Department of Anesthesiology, Perioperative and Pain Medicine, Laboratory for Tissue Engineering and Regenerative Medicine, Brigham and Women's Hospital, 75 Francis St., Boston, MA 02115, USA.
Kojima
Koji
K
Vacanti
Charles A
CA
Kodama
Shohta
S
D016475:3T3 Cells / Q000166:cytology* / Q000187:drug effects; D000818:Animals; D001973:Bromodeoxyuridine / Q000493:pharmacokinetics; D002452:Cell Count; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects*; D018844:Cyclin-Dependent Kinases / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D008012:Lidocaine / Q000494:pharmacology*; D051379:Mice; D012313:RNA / Q000235:genetics; D054462:p21-Activated Kinases / Q000235:genetics*
D012313:RNA; D008012:Lidocaine; C515905:Pak1 protein, mouse; D054462:p21-Activated Kinases; D018844:Cyclin-Dependent Kinases; D001973:Bromodeoxyuridine